1. Field of the Invention
The present invention relates to a composition for inhibiting HIV activity comprising the extract of Paecilomyces sp. (Tochu-kaso) J300. More particularly, the present invention relates to a pharmaceutical composition for inhibiting HIV activity comprising 3-[5-(methoxy-ethyl)-3,6-dioxo-piperazine-2-yl]propionic acid represented by Formula 1, 4-methyl-2-[(pyrolidine-2-carbonyl)-2-amino]pentanoic acid represented by Formula 2 or derivatives thereof that can be extracted from Paecilomyces sp. (Tochu-kaso) J300; and to a and food composition containing the same.

2. Description of the Related Arts
From the ancient time, the tochu-kaso was regarded as miraculous medicine, and especially, Chinese Emperor Jinsihwang took the tochu-kaso as perpetual youth and longevity. Further, the recent reports disclose that tochu-kaso has the effect of anti-exhaustion, immunity-increasing and anti-tumor activities, etc. (Natural Products Research Institute of Seoul National University). The various effects of the tochu-kaso are thought to originate from the various components of the tochu-kaso and its direct or indirect relation to the homeostasis of the human body.
There are 300 kinds of tochu-kasos are known in the world, but the size and the number of the population is small, and that, it is difficult to obtain them commercially and even samples for research are not sufficient. Therefore, Rural Development Administration of Korea has invented a the method for cultivating the tochu-kaso by inoculating the spores of the tochu-kaso into a larva of silkworm. In the year of 2000, the method of cultivation is discovered for 5 kinds of tochu-kasos including Paecilomyces japonica. Paecilomyces sp. J300 (Tochu-kaso J300) is obtained by inoculating the spores of the Paecilomyces sp. J300 into a larva of silkworm, and the method for cultivating the tochu-kaso J300 is disclosed Korean Patent No 187,897 and U.S. Pat. No. 5,939,310.
According to UNAIDS and WHO, there are more than thirty million (30,000,000) people who are infected by HIV in the year of 1997 throughout the world. The number of people newly infected by HIV in the year 1997 is about 5,800,000 and 590,000 of those are children. With this increase, the number of people infected by HIV seems to be forty million (40,000,000) in the year of 2000. More than 90% of the infected are living in the developing countries, and most of the infected do not know the fact that they are infected, which is a serious problem.
Therefore, eliminating AIDS is an international issue, and a new medicine or medical composition to eliminate AIDS without side effects are eagerly being sought.
After the first sufferer of AIDS is reported in 1981, HIV, the AIDS Virus was separated and identified from a sufferer of AIDS in 1984, and HIV is convinced to be main cause of the AIDS. Therefore, to treat AIDS, searching for anti-HIV material is widely performed. As the result, AZT, the most famous AIDS treatment, was invented in 1987, and at the time of February 1999, there are 13 kinds of drugs approved by FDA and sold as AIDS treatment. However, the conventional treatment for AIDS has the problems that resistant virus is generated and side effect is observed, and therefore, there are limitations in the treatments.
To overcome or reduce the limitations, many pharmaceutical companies and research centers are searching for new and developed AIDS treatments.
HIV (Human Immunodeficiency Virus) was disclosed to be cause of AIDS by being separated and identified from a sufferer of AIDS in 1984. At the beginning of the disclosure, HIV infection was misunderstood to be restricted only to homosexual or some under developed countries, but now, it is regarded as the worldwide prevalent infectious disease and a serious social problem.
HIV is classified in the retrovirus, more specifically, in the lentivirus group. The size of the HIV is about 10 micron and the outer surface is covered with phospholipid like other cells, and two virus genomes of RNA are protected by capsid (core protein) therein. These HIV genomes consist of ten (10) units of genes, which is a lot of genes compared with the size of total genome.
HIV infection is accomplished by the conjugation of the envelop protein (gp120) that exists on the surface of the virus and receptor that exists on the surface of the target cell. The above receptor is a kind of surface protein molecule of cell called CD4 antibody, and CD4 T cell (helper T cell) or macrophage is main target of the HIV because they have a lot of CD4 antibodies on the surface thereof. By the conjugation of the virus and the cell, the phospholipid envelop of the virus is infused into the cell surface, and virus genome and nucleus protein are flowed into the cell. At this moment, the virus genome is reverse transcribed into DNA from RNA by the reverse transcriptase of the virus, then transported into the nucleus of the cell and inserted into the genome of the host cell. This procedure is one of the characteristic features of the retrovirus. The HIV conceals itself in the most safe region of the host cell, and is supplied with all the materials and nutritions needed for growth. Further, the HIV suppresses or accelerates its growth according to the surroundings and conditions thereof to protect itself from the outer antibody etc.
AIDS viruses are divided tow main groups, that are, HIV-1 and HIV-2. The HIV-1 is generally called as ‘AIDS virus’ because it is generally found various sufferers throughout the world including Korea. HIV-2 that is found from the sufferer s of the West Africa has only 55% of homologies with the HIV-1 in the base sequence, and is more similar to SIV (Simian Immunodeficiency Virus) that is monkey's AIDS virus. The toxicity of the HIV-2 is known to be weaker than HIV-1.
HIV has much variety in the viewpoints of biology as well as genetics. Base sequences of the HIV obtained from the sufferers are different each other. And, even when the base sequence is obtained from one sufferer, it is different according the state of progress. Further, when the viruses are obtained with a uniform time intervals, the base sequence is different according to the tissues. These various base sequence relates to the biological characteristics of the virus very much. The different viruses having different base sequence have different affinity of infection to a certain cell, rate of proliferation, generation of virus, toxicity to a cell, generation rate of multinucleate giant cell, latent period and activity period, sensitivity to the neutralizing antibody, etc.
According to examination result for the relation betweens these various biological characteristics and the onset of the AIDS, the viruses that obtained from the initially infected sufferer do not form a multinucleate giant cell (NSI: Nonsyncytia-Inducing), but are mainly infected via macrophage. However, as the AIDS proceeds to the terminal state, the SI (Syncytia-Inducing) increase, and the virus changes to be a virus that is likely to infected by helper T cell instead of macrophage. This suggests that the biological characteristics of the HIV and onset of the disease have relations each other.
When one week is passed after HIV infection, proliferation of virus increases, and that, the virus is easily found in the blood of the sufferer. This stage is called as viremia. After 1 or 2 week later, the number of virus dramatically decreases under the level not to be separated. This latent period is maintained for a long time, then the proliferation of virus becomes active to develop for AIDS and becomes the state of viremia again. However, a recent research based on the study of chain polymerization with polymerase reported that virus is continuously generated in the latent period.
The number of CD4 cell dramatically decreases in the first viremia, and when the proliferation of the virus decreases the number of CD4 cell is recovered to its normal state. (The number of CD4 cell of the healthy adult is 500-1000 CD4 cells/mm3). When the number of CD4 cells decrease under 200 cells/mm3, it develops as ARC (AIDS-related Complex) or AIDS. Because the rate of opportunistic infection increases for the AIDS sufferers, they mainly die from Pneumocysitis carinii.
The periods that the number of HIV dramatically decreases and that the number of CD8 cells increases coincide, and CD8 T cell is known as to inhibit the growth of cells or to kill the cells infected by virus. Therefore, CD8 cell shows an important immunity effect for the initially infected viruses. Antibody is generated after the virus decreases. CD 8 cell and antibody are detected from the beginning of the infection to the onset of AIDS, but their activities become disappeared and in some cases they promote the virus infection. The reason why the immunity system that showed anti-virus activity at the beginning of the infection loses its activity is a subject to be solved. Because AIDS outbreaks only in the human body, the research is limited and the understanding of this disease is very rudimentary.
The researchers agree that the cause of the Immune Deficiency is the decrease of the CD4 cell, however, they have different opinions regarding the mechanism that HIV decreases the CD4 cell. For the explanation of the CD4 cell decrease, there are many theories such as generation of multinucleate giant cell, stimulation of virus DNA that is not inserted, the influence by the constitution of the host cell membrane, programmed abandonment of death of the cell, secretion of toxic material from the infected cell, cell destruction by auto-immunity, et al. However, nothing was proved in vivo. Further, many researches have been made to figure out the symptom of HIV disease by using monkey and monkey's AIDS virus, SIV, but no significant discovery was not accomplished.
In the meantime, AZT (Zidovudine), regarded as the representative of AIDS treatment, is a drug that inhibits the activity of the HIV reverse transcriptase and its effects are widely studied. AZT improves the condition of the sufferer when used at the initial state of infection, however it cannot lengthen the lifetime of the sufferer. Further, it has a toxic influence to the marrow and resistant virus is generated when used for a long time. Therefore, it has a limitation for the AIDS treatment. As for DDI, DDC, d4T etc. that are approved by FDA for the treatment of AIDS, resistant viruses have been generated, but they are not so toxic as AZT. Recently, a theory suggesting that using the above drugs together can reduce or inhibit the toxicity and generation of resistant virus as well as can increase the therapeutic effects is reported. To clarify the suggestion, a lot of research institutes have performed clinical tests, and positive results were obtained.
In addition to the above research, there are a lot of smart and novel methods for inhibiting the growth of AIDS virus are invented and testified. For example, a method preventing the virus from accessing to cells, a method selectively destructing virus-infected cells, a therapeutic method of using a drug for inhibiting the important activity of enzyme for the growth of virus (such as protease, integrase, tat inhibitor, rev inhibitor) or cytokine, a method of inserting CD8 cell, gene therapeutics etc. are suggested.
There was significant improvement in the research of the HIV vaccine. Various methods such as using died virus or attenuated virus, using subunit vaccine by genetic engineering, using anti-idio type antibody, injecting DNA gene etc. were developed. The problem of developing vaccine is that the virus is too various. For example, after vaccination, when using the original virus used for above vaccine the disease is contracted, the growth of virus is suppressed by the antibody; however, when the disease is contracted by using newly infected cell or virus, no suppression was observed. It is a subject for the inventors to overcome the variety of the viruses. In addition to the vaccine using body fluid, the vaccines using cell immune method with CD8 cell or mucosal immunity should be developed, and more effective results are expected by mixing the vaccines.
Under these circumstances, the present inventors studied to develop a new AIDS treatment without side effects and generation of the resistant virus.
As the result, the inventors found that the extract of J300 Tochu-kaso contains anti HIV activity, and analyzed the active fraction to find out active component, further, the active component was synthesized by chemically to provide novel AIDS treatment with low toxicity and side effect.